Human milk fortifier

ABSTRACT

A human milk fortifier composition comprising one or more human milk oligosaccharides. Said human milk fortifier composition may be tailored to fortify the breast milk of a primiparous woman.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a human milk fortifier composition, more specifically to a human milk fortifier composition comprising human milk oligosaccharides. In particular the present invention relates to a human milk fortifier composition specifically tailored to fortify the breastmilk of a primiparous woman, and for consumption by an infant or child born who is the offspring of a primiparous mother. The invention furthermore relates to the use of said human milk fortifier composition.

BACKGROUND OF THE INVENTION

Infants who are the offspring of primiparous woman are considered to be at an increased risk of suffering from a variety of health complaints in infancy, childhood, and later life. Infants born of primiparous mothers are more likely to be smaller and/or thinner at birth, and so at an increased risk of suffering from conditions associated with being small and/or underweight, and at the same time are also more likely to be larger children and suffer from conditions associated with being larger e.g. being overweight or obese—this is because of dramatic catch-up gains in weight and length that overcompensate for their initial size deficit.

The reasons for the increased health risks seen in infants who are the offspring of primiparous mothers is not clear. However, there is a need to identify factors that may contribute to this risk and to address them.

The inventors have now identified a factor that they believe may contribute to this risk. In particular the inventors have found that the concentration of one or more human milk oligosaccharides (herein after “HMOs”) found in human breast milk (hereinafter “HM”) produced by primiparous mothers, may differ from the concentration found in HM produced by multiparous mothers. More particularly the inventors have found that the concentration of an HMO found in HM produced by primiparous mothers may be lower than the concentration of the same HMO found in HM produced by multiparous mothers.

HMOs are, collectively, the third largest solid constituents in human milk, and a variety of benefits have been associated with them, in consequence, an optimal intake of these compounds in infancy and childhood is believed to be necessary to ensure optimum health and development. HMOs have for example been linked to a variety of biological functions including the establishment of gut microbiota. Some HMOs have also been linked to growth and body composition.

Accordingly, there is need for milk fortifiers comprising one or more HMO that can be used to fortify HM produced by primiparous mothers, and to optimise the intake of one or more HMO in infants consuming HM from a primiparous mother.

SUMMARY OF THE INVENTION

The invention is set out in the claims and in the detailed description included herein. The inventors have found that the concentration of an HMO found in HM produced by multiparous mothers may be higher than the concentration of the same HMO found in HM produced by primiparous mothers. In light of this finding, the inventors have developed a human milk fortifier composition comprising one or more HMOs.

Said human milk fortifier may be tailored to fortify the breast milk of a primiparous women.

The one or more HMO may be a sialylated oligosaccharide, a fucosylated oligosaccharide, an N-acetylated oligosaccharide, or any combination thereof. The one or more HMO may for example be selected from the group consisting of; 2′-Fucosyllactose, 3′-Fucosyllactose, 6′Galactosyllactose, Blood group A Tetrasaccharide, Difucosyllacto-N-Hexose-A, Lacto-N-Difucosylhexose, Lacto-N-Fucosylpentaose-I, Lacto-N-Fucosylpentaose-II, Lacto-N-Fucosylpentaose-III, Lacto-N-Fucosylpentaose-V, Lacto-N-hexaose A, Lacto-N-hexaose B, Lacto-N-Neodifucosylhexose, Lacto-N-Neofucosylpentaose, Lacto-N-Neotetraose, Lacto-N-Tetraose, Lactodifucosyllactose, Sialyllacto-N-Tetraose a, Sialyllacto-N-Tetraose b, and any combination thereof.

It may be particularly beneficial if the HMO is selected from the group consisting of; 3′-Fucosyllactose, Lacto-N-Fucosylpentaose, and any combination thereof.

The human milk fortifier composition may comprise an HMO in a range of 0.1 to 10000 mg/L.

The human milk fortifier may be specifically tailored for up an infant of an age selected from the group consisting of; up to 4 months of age, up to 3 months of age, up to 2 months of age, up to 1 months of age, up to 2 weeks of age, and up to 1 week of age. It may for example be specifically tailored to supplement breastmilk produced for up an infant of up to one month of age or up to 2 weeks of age. The infant may be an infant of a primiaparous woman.

The human milk fortifier may further comprise one or more ingredient selected from the group consisting of vitamins, minerals, protein, carbohydrates, and probiotics.

Further provided is a method of preparing a human milk fortifier composition tailored to fortify the breast milk of a primiparous women, said method comprising the steps of: measuring out an appropriate amount of a human milk fortifier composition and mixing it with a diluent and/or additive.

Also provided is a human milk fortifier as defined herein, for use in fortifying human breast milk and in particular human breastmilk from a primiparous woman.

The human milk fortifier as defined herein may to provide an optimised amount of one or more HMO to an infant. The infant may be selected from the group consisting of: preterm infants and term infants. The infant may be an infant who is the offspring of a primiparous mother.

Also provided is a nutritional system comprising:

a. a human milk fortifier composition tailored to fortify the breast milk of a primiparous women, and b. A human milk fortifier composition, wherein, said human milk fortifier composition tailored to fortify the breast milk of a primiparous women comprises one or more HMO in a concentration higher than in the human milk fortifier composition. The method may also comprise the step of determining whether the woman is a primiaparous woman.

DRAWINGS

FIG. 1 is a graphical representation of the 2′-fucosyllactose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 2 is a graphical representation of the 3-Fucosyllactose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 3 is a graphical representation of the 6′Galactosyllactose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 4 is a graphical representation of the Blood group A Tetrasaccharide concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 5 is a graphical representation of the Difucosyllacto-N-Hexose-A concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 6 is a graphical representation of the Lacto-N-Difucosylhexose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 7 is a graphical representation of the Lacto-N-Fucosylpentaose-I concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 8 is a graphical representation of the Lacto-N-Fucosylpentaose-II concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 9 is a graphical representation of the Lacto-N-Fucosylpentaose-III concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 10 is a graphical representation of the Lacto-N-Fucosylpentaose-V concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 11 is a graphical representation of the Lacto-N-hexaose A concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 12 is a graphical representation of the Lacto-N-hexaose B concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 13 is a graphical representation of the Lacto-N-Neodifucosylhexose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 14 is a graphical representation of the Lacto-N-Neofucosylpentaose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 15 is a graphical representation of the Lacto-N-Neotetraose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 16 is a graphical representation of the Lacto-N-Tetraose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 17 is a graphical representation of the Lactodifucosyllactose concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 18 is a graphical representation of the Sialyllacto-N-Tetraose a concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

FIG. 19 is a graphical representation of the Sialyllacto-N-Tetraose b concentration found in HM by delivery mode at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum.

DETAILED DESCRIPTION

In a first aspect of the present invention there is provided a human milk fortifier composition comprising one or more HMO.

The term “human milk fortifier composition” as used herein, refers to a nutritional composition for use in combination and in admixture with human breast milk. Unless otherwise specified, the term “human milk fortifier composition” specifically excludes conventional infant formulas that provide the sole or primary source of infant nutrition and that are not typically combined and admixed with human milk to supplement human milk feedings.

The term “fortifier” refers to a composition which comprises one or more nutrients having a nutritional benefit for infants, both preterm infants and term infants. The fortifier according to the present invention is rich in HMOs and may therefore be considered as an HMO fortifier, HMO supplement or the like.

In an embodiment of the invention, the human milk fortifier composition is tailored/adapted to fortify the breast milk of a primiparous women. A human milk fortifier, as disclosed herein, may be considered as specifically tailored/adapted to fortify the breast milk of a woman who is primaprous if it comprises one or more HMO as described herein. Said human milk fortifier may, for example, comprise said one or more HMO in an amount sufficient to address the deficiency of one or more HMO identified in the human breast milk of primiaparous mothers in comparison to multiparous mothers. A sufficient amount of an HMO may for example be an amount equal to or greater than an amount that an infant born to a multiparous woman would receive, or may for example, be any amount that is equal to or higher than the difference found in the concentration e.g. averages, in human milk produced by primparous women and multiparous women. Said human milk fortifier composition may be a parity specific human milk fortifier i.e. a milk fortifier sold specifically for use in primiparous women e.g. marketed as a being for use to fortify the breastmilk of primiparous women

The term “multiparous” as used herein refers to a woman who has given birth more than once or has more than 1 child.

The term “primiparous” as used herein refers to a woman who has given birth once, or has only 1 child.

The term “infant” as used herein, refers to humans of less than about 1 year of age. The term includes preterm infants, premature infants, small for gestational age (SGA) infants and/or infant with low birth weight (LBW).

The terms “preterm infants” or “premature infants” as used herein, refer to infants who were not born at term. Generally they refers to infants born alive prior to 37 weeks of gestation/pregnancy.

The term “small for gestational age infant” as used herein, refers to an infant who is smaller in size than normal for their gestational age at birth, most commonly defined as a weight below the 10th percentile for the gestational age. In some embodiments, SGA may be associated with intrauterine growth restriction (IUGR), which refers to a condition in which a foetus is unable to achieve its potential size.

The term “low birth weight infants” as used herein refers to an infant that has a body weight under 2500 g at birth. It therefore encompasses:

-   -   infants who have/had a body weight from 1800 to 2500 g at birth         (usually called “low birth weight” or LBW)     -   infants who have/had a body weight from 1000 to 1800 g at birth         (called “very low birth weight” or VLBW)     -   infants who have/had a body weight under 1000 g at birth (called         “extremely low birth weight” or ELBW) Infants or young children         with low birth weight may or may not be preterm, and similarly,         infants or young children who were small for gestational age may         or may not be preterm.

The term “child” as used herein, refers to humans from about 1 to about 7 year of age, for example, between 1 and 3 years of age.

The human milk fortifier composition of the invention may comprise any type of HMO.

In an embodiment of the present invention the human milk fortifier comprise a HMO selected from the group consisting of a sialylated oligosaccharide, a fucosylated oligosaccharide, an N-acetylated oligosaccharide, or any combination of the foregoing.

The term “sialylated oligosaccharide” as used herein refers to an oligosaccharide having a sialic acid (such as N-acetylneuraminic acid and/or N-glycolylneuraminic acid) residue.

The term “N-acetylated” oligosaccharide as used herein refers to an oligosaccharide having at least one hexose carrying an N-acetyl residue.

The term “fucosylated oligosaccharide” as used herein refers to an oligosaccharide having a fucose residue.

In a more specific embodiment the human milk fortifier composition of the invention comprises an HMO selected from the group consisting of: 2′-Fucosyllactose, 3-Fucosyllactose, 6′Galactosyllactose, Blood group A Tetrasaccharide, Difucosyllacto-N-Hexose-A, Lacto-N-Difucosylhexose, Lacto-N-Fucosylpentaose-I, Lacto-N-Fucosylpentaose-II, Lacto-N-Fucosylpentaose-III, Lacto-N-Fucosylpentaose-V, Lacto-N-hexaose A, Lacto-N-hexaose B, Lacto-N-Neodifucosylhexose, Lacto-N-N eofucosylpentaose, Lacto-N-Neotetraose, Lacto-N-Tetraose, Lactodifucosyllactose, Sialyllacto-N-Tetraose a, Sialyllacto-N-Tetraose b, and any combination thereof.

In an even more specific embodiment the human milk fortifier composition of the invention comprises an HMO selected from the group consisting of: 3-Fucosyllactose, Lacto-N-Fucosylpentaose II, 2′-Fucosyllactose and any combination thereof.

The human milk fortifier composition of the invention may comprise an HMO in any concentration.

In particular the human milk fortifier composition may comprise one an HMO in a concentration of 0.1 to 10000 ring/L e.g. 0.1 to 6000 mg/L e.g. 0.1 to 350 mg/L.

The concentrations listed herein may refer to a concentration after a composition has been reconstituted or mixed with water or milk.

The human milk fortifier composition of the invention may for example comprise one or more of the HMOs listed in table I in a concentration range listed in table I.

TABLE I HMO Concentration Range mg/L 2′-Fucosyllactose   9-8500 3-Fucosyllactose   1-3600 6′Galactosyllactose 0.09-250  Blood group A Tetrasaccharide   1-1000 Difucosyllacto-N-Hexose-A  0.4-1100 Lacto-N-Difucosylhexose   20-3000 Lacto-N-Fucosylpentaose-I  0.4-4300 Lacto-N-Fucosylpentaose-II   32-3600 Lacto-N-Fucosylpentaose-III  13-800 Lacto-N-Fucosylpentaose-V 1.5-500 Lacto-N-hexaose A 0.15-200  Lacto-N-hexaose B 0.3-120 Lacto-N-Neodifucosylhexose 0.3-330 Lacto-N-Neofucosylpentaose 0.1-65  Lacto-N-Neotetraose 0.1-700 Lacto-N-Tetraose   4-3400 Lactodifucosyllactose   2-3300 Sialyllacto-N-Tetraose a  1.5-1900 Sialyllacto-N-Tetraose b 0.7-180

In an embodiment of the present invention the human milk fortifier composition of the invention may comprise one or more of the HMOs listed in table II in the concentration range listed in table II.

TABLE II HMO Concentration Range mg/L 2′-Fucosyllactose  9-350 3-Fucosyllactose 51-230 6′Galactosyllactose 0.09-11   Blood group A Tetrasaccharide 1.5-22   Difucosyllacto-N-Hexose-A 0.4-9   Lacto-N-Difucosylhexose 20-160 Lacto-N-Fucosylpentaose-I 0.4-75   Lacto-N-Fucosylpentaose-II 14-190 Lacto-N-Fucosylpentaose-III 13-16  Lacto-N-Fucosylpentaose-V 1.5-22   Lacto-N-hexaose A 2-4  Lacto-N-hexaose B  1-4.5 Lacto-N-Neodifucosylhexose 1.1-9   Lacto-N-Neofucosylpentaose 1.8-6   Lacto-N-Neotetraose 0.1-6   Lacto-N-Tetraose 23-125 Lactodifucosyllactose 2-38 Sialyllacto-N-Tetraose a 1.5-36   Sialyllacto-N-Tetraose b 0.7-11  

The human milk fortifier of the invention may be tailored to fortify breastmilk produced for an infant or child of any age e.g. any age born to a primparous mother

In an embodiment of the present invention the human milk fortifier composition is tailored/adapted to fortify breastmilk produced for an infant of an age selected from the group consisting of; up to 4 months of age, up to 3 months of age, up to 2 months of age, up to 1 months of age, up to 2 weeks of age, up to 1 week of age. For example the human milk fortifier composition may be tailored/adapted to fortify breastmilk produced for an infant up to 1 month of age e.g. up to 2 weeks of age. The infant may be born to a primparous mother.

In an embodiment of the present invention the human milk fortifier is tailored/adapted for an infant of up to 1 month of age e.g. an infant up to 2 weeks of age, or an infant up to 1 week of age and said composition comprises one or more HMO selected from the group consisting of 3-Fucosyllactose, Lacto-N-Fucosylpentaose II, 2′-Fucosyllactose, and any combination thereof. In a more specific embodiment said HMOs, if present in said human milk fortifier tailored/adapted for an infant of up to 1 month of age, may be present in a concentration range as set out in table III.

TABLE III HMO Concentration Range mg/L 3-Fucosyllactose 51-230 Lacto-N-Fucosylpentaose II 14-190 2′-Fucosyllactose  9-350

The human milk fortifier composition of the invention can also comprise any other ingredients or excipients known to be employed in human milk fortifier compositions.

Non limiting examples of such ingredients include: proteins, amino acids, carbohydrates, lipids, prebiotics or probiotics, essential fatty acids, nucleotides, nucleosides, vitamins, minerals and other micronutrients.

In an embodiment of the invention the human milk fortifier composition further comprises one or more ingredient selected from the group consisting of vitamins, minerals, protein, carbohydrates, and probiotics.

Non limiting examples of proteins include: casein, alpha-lactalbumin, whey, soy protein, rice protein, corn protein, oat protein, barley protein, wheat protein, rye protein, pea protein, egg protein, sunflower seed protein, potato protein, fish protein, meat protein, lactoferrin, serum albumin, immunoglobins, and combinations thereof.

Non limiting examples of amino acids include leucine, threonine, tyrosine, Isoleucine, arginine, alanine, histidine, isoleucine, proline, valine, cysteine, glutamine, glutamic acid, glycine, serine, arginine, lysine, methionine, phenylalanine, tryptophane, asparagine, aspartic acid, and combinations thereof.

Non limiting examples of carbohydrates include lactose, saccharose, maltodexirin, starch, and combinations thereof.

Non limiting examples of lipids include: palm olein, high oleic sunflower oil, high oleic safflower oil, canola oil, fish oil, coconut oil, bovine milk fat, and combinations thereof.

Non limiting examples of essential fatty acids include: linoleic acid (LA), a-linolenic acid (ALA) and polyunsaturated fatty acids (PUFAs). The gender specific synthetic nutritional compositions of the invention may further contain gangliosides monosialoganglioside-3 (GM3) and disialogangliosides 3 (GD3), phospholipids such as sphingomyelin, phospholipids phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and combinations thereof.

None limiting examples of prebiotics include: oligosaccharides (other than HMOs) optionally containing fructose, galactose, mannose; dietary fibers, in particular soluble fibers, soy fibers; inulin; and combinations thereof. Preferred prebiotics are fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS), isomalto-oligosaccharides (IMO), xylo-oligosaccharides (XOS), arabino-xylo oligosaccharides (AXOS), mannan-oligosaccharides (MOS), oligosaccharides of soy, glycosylsucrose (GS), lactosucrose (LS), lactulose (LA), palatinose-oligosaccharides (PAO), malto-oligosaccharides, gums and/or hydrolysates thereof, pectins and/or hydrolysates thereof, and combinations of the foregoing.

Non limiting examples of probiotics include: Bifidobacterium, Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Kluyveromyces, Saccharoymces, Candida, in particular selected from the group consisting of Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus lactis, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Saccharomyces cerevisiae, Saccharomyces boulardii or mixtures thereof, preferably selected from the group consisting of Bifidobacterium longum NCC3001 (ATCC BAA-999), Bifidobacterium longum NCC2705 (CNCM I-2618), Bifidobacterium longum NCC490 (CNCM I-2170), Bifidobacterium lactis NCC2818 (CNCM I-3446), Bifidobacterium breve strain A, Lactobacillus paracasei NCC2461 (CNCM I-2116), Lactobacillus johnsonii NCC533 (CNCM I-1225), Lactobacillus rhamnosus GG (ATCC53103), Lactobacillus rhamnosus NCC4007 (CGMCC 1.3724), Enterococcus faecium SF 68 (NCC2768; NCIMB10415), and combinations thereof.

Non limiting examples of Nucleotides include: cytidine monophosphate (CMP), uridine monophosphate (UMP), adenosine monophosphate (AMP), guanosine monophosphate (GMP), and combinations thereof.

Non limiting examples of vitamins and minerals include: vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin Bi2, vitamin E. vitamin K. vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorous, iodine, iron, magnesium, copper, zinc, manganese, chloride, potassium, sodium, selenium, chromium, molybdenum, taurine, L-carnitine, and combinations thereof. Minerals are usually added in salt form.

Other suitable and desirable ingredients of human milk fortifier compositions, that may be employed in the human milk fortifier compositions of the invention, are described in guidelines issued by the Codex Alimentarius.

The human milk fortifier composition of the invention may be prepared in any way known in the art to prepare human milk fortifier compositions. It is well within the purview of the skilled person to decide on a method depending on the type of human milk fortifier in question e.g. powder or liquid. An exemplary method for preparing a human milk fortifier in accordance with the invention is set out below.

A human milk fortifier may be prepared, for example, by blending together lipid, protein, HMOs, and carbohydrate in appropriate proportions. If used, emulsifiers may be included in the blend at this stage. The vitamins and minerals may be added at this stage but are usually added later to avoid thermal degradation. Any lipophilic vitamins, such as vitamin A, D, E and K, and emulsifiers may be dissolved into the fat source prior to blending. Water, preferably water which has been subjected to reverse osmosis, may then be mixed in to a liquid mixture.

The liquid mixture may then be thermally treated to reduce bacterial loads. For example the liquid mixture may be rapidly heated to a temperature on the range of about 80° C. to about 110° C. for about 5 seconds to about 5 minutes. This may be carried out by steam injection or by heat exchanger, for example a plate heat exchanger.

The liquid mixture may then be cooled to about 60° C. to about 85° C., for example by flash cooling. The liquid mixture may then be homogenised, for example in two stages at about 7 MPa to about 40 MPa in the first stage and about 2 MPa to about 14 MPa in the second stage. The homogenised mixture may then be further cooled and any heat sensitive components, such as vitamins and minerals may be added. The pH of the homogenised mixture is conveniently standardised at this point.

The homogenized liquid mixture is then filled into suitable containers, preferably aseptically. However, the liquid composition may also be reported in the container. Suitable apparatus for carrying out filling of this nature is commercially available.

A human milk fortifier composition specifically tailored/adapted to fortify the breast milk of a primiparous women may be prepared from a human milk fortifier composition e.g. a human milk fortifier composition not specifically tailored to fortify the breast milk of a woman of a particularly parity e.g. primiparous or multiparous.

Accordingly, in another aspect of the present invention there is provided a method of preparing a human milk fortifier composition tailored to fortify the breast milk of a primiparous women, said method comprising the steps of: measuring out an appropriate amount of a human milk fortifier composition e.g. a human milk fortifier composition not specifically tailored to fortify the breast milk of a woman of a particular parity, and mixing it with an additive and/or a diluent e.g. one or more HMOs and/or water, so as to arrive at a human milk fortifier composition tailored to fortify the breast milk of a primiparous woman in accordance with the invention.

The additive may be a one or more HMO e.g. one or more HMO in a concentration such, that that when the additive is mixed with a human milk fortifier composition, and optionally a diluent, the resulting mixture is a human milk fortifier tailored to fortify the breast milk of a primiparous women, in accordance with the invention.

The additive may be a parity specific additive e.g. an additive marketed as specifically being for use by primiparous women.

In another aspect of the present invention there is provided a human milk fortifier in accordance with the invention, for use in fortifying human breast milk.

In an embodiment the human breastmilk is breastmilk from primiparous women.

In another aspect of the present invention there is provided a human milk fortifier composition in accordance with the invention, for use to provide an optimised amount of and/or to prevent a sub-optimal intake of one or more HMO to an infant or child who is the offspring of a primiparous mother.

An optimised amount of one or more HMO would be an amount equal to or greater than an amount e.g. the average amount, that an infant born to a multiparous woman would be considered to receive e.g. an amount of an HMO set out in table I, II or III included herein.

In another aspect of the present invention there is provided a human milk fortifier composition in accordance with the invention, for use in optimising the health and development and/or preventing the sub-optimal health and development e.g. growth and development of an infant or child who is the offspring of a primiparous mother.

The human milk fortifier compositions of the invention may not only optimise the health and development of an infant or child who is the offspring of a primiparous mother in the short term, but may also do so in the long term.

In another aspect of the present invention there is provided a human milk fortifier composition in accordance with the invention, for use in optimising the gut microbiota and/or preventing sub-optimal gut microbiota in an infant or child who is the offspring of a primiparous mother. HMOs are known to be important for the establishment of gut microbiota and therefore an optimal supply of HMOs may lead to an optimised gut microbiota.

In another aspect of the present invention there is provided the use of a human milk fortifier composition in accordance with the invention, in the manufacture of a composition for use in optimising the gut microbiota and/or health and development of an infant or child who is the offspring of a primiparous mother.

A non-optimal gut microbiota may be one showing presence of one or several pathobionts and/or opportunistic pathobionts and/or their toxins and/or virulence factors and/or antibiotic resistence genes. An optimal gut microbiota may be one not showing presence of one or several pathobionts and/or opportunistic pathobionts and/or their toxins and/or virulence factors and/or antibiotic resistence genes.

The human milk fortifier compositions of the invention may not only optimise gut microbiota composition and/or health and development short term, but may also do so in the long term.

Long term effects may only be evident in months or years e.g. 6 months, 9 months, 12 months, 5 years, 10 years, or 20 years

In another aspect of the present invention there is provided the use of a human milk fortifier composition in accordance with the invention, to fortify human breast milk and to improve/prevent sub-optimal breastmilk quality wherein, said breastmilk is from a primiparous women.

The quality of breastmilk in a primparous woman may be considered sub-optimal if it comprises one or more HMO in a concentration less than that found in breastmilk from a multiparous woman e.g. in a concentration less than the average found in multiparous women.

In another aspect of the present invention there is provided the use of a human milk fortifier according to the invention in optimising and/or preventing the sub-optimal health and development and/or the gut flora composition in an infant or child who is the offspring of a primiparous mother.

As stated previously, some HMOs have also been linked to growth and body composition. In particular the intake of some HMOs have been positively linked to weight, lean mass and fat mass. In particular Lacto-N-Fucosylpentaose-II has been linked to increased size and in particular increased fat mass.

In another aspect of the present invention there is provided a human milk fortifier according to the invention for use in optimising and/or preventing the sub-optimal growth of an infant or child who is the offspring of a primiparous mother. In an embodiment the human milk fortifier according to the invention comprises Lacto-N-Lacto-N-Fucosylpentaose-II. In a more specific embodiment the human milk fortifier is tailored/adapted to fortify the human breast milk produced for an infant of up to 4 months of age, more specifically up to 1 month of age e.g. up to 2 weeks of age.

An infants or child's growth and development may be optimised if for example said infants or child's weight and length is within growth curves developed by the WHO.

Optimized growth refers to the criteria set by the WHO and they are depicted as charts of the growth of breastfed infants with the appropriate age- and sex-specific weight, length and head circumference. The WHO growth charts are international standards that show how healthy infants should grow, describing the growth of infants living in environments believed to support optimal growth.

If an infant or child's weight and length is within the growth curves developed by the WHO, and he/she has an appropriate distribution of lean and fat mass, it may be assumed that this child/infant is growing and developing in a healthy way, and may have a lower risk for later adverse metabolic outcomes.

Health and development and/or gut flora composition and/or growth may be optimised short term or long term.

A human milk fortifier tailored to fortify the breastmilk of a primiparous woman may be included in a nutritional system.

The term “nutritional system” as used herein refers to a collection of more than one synthetic nutritional compositions advertised or sold as part of the same product range e.g. a collection of human milk fortifiers and/or infant formulas sold under the same brand and adapted/tailored to the nutritional needs of infants born to different parity mothers e.g. primiparous or multiparous mothers. The synthetic nutritional compositions making up the nutritional system may be packaged individually e.g. in capsules or boxes. Said packages can be sold individually, grouped together e.g. wrapped by plastic film or combined in a box, or in a combination of these two ways. The nutritional system may also comprise synthetic nutritional compositions for children older than 12 months.

In a further aspect of the present invention there is provided a nutritional system comprising:

-   -   a. a human milk fortifier composition tailored to fortify the         breast milk of a primiparous women, in accordance with the         invention, and     -   b. a human milk fortifier composition e.g. a human milk         fortifier composition not specifically tailored to fortify the         breast milk of a woman of a specific parity or specifically         tailored to fortify the offspring of a multiparous woman,

wherein,

said human milk fortifier composition tailored to fortify the breast milk of a primiparous women comprises one or more HMO in a concentration higher than in the human milk fortifier composition e.g. human milk fortifier composition not specifically tailored to fortify the breast milk of a woman of a specific parity.

The concentration of one or HMO in the human milk fortifier tailored for a primiparous woman may be higher by any amount.

In an embodiment the human milk fortifier composition tailored for a primiparous woman comprises one or more HMO listed in table II in a higher amount. The higher amount may be an amount within the range given in table II for the HMO in question.

In a more specific embodiment the human milk fortifier composition tailored for a multiparous woman comprises one or more HMO listed in table III in a higher amount. The higher amount may be an amount within the range given in table III for the HMO in question.

It should be appreciated that all features of the present invention disclosed herein can be freely combined and that variations and modifications may be made without departing from the scope of the invention as defined in the claims. Furthermore, where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred to in this specification.

There now follows a series of non-limiting examples that serve to illustrate the invention.

EXAMPLES Example 1 Longitudinal Clinical Trial:

The present inventors designed a longitudinal clinical trial with lactating mothers with milk sampling at 2 days (V1), 17 days (V2), 30 days (V3) 60 days (V4), 90 days (V5), and 120 days (V6) postpartum. The milk samples were quantitatively analyzed for HMOs.

The data presented here is from a multi-center, exploratory study with the primary objective of characterizing key nutrient components in human breast milk. Healthy women of any ethnicity having decided to exclusively breast-feed their new born infant from birth to 4 months of infant's age were recruited during the last 3 months of pregnancy, and their infants were followed up until 4 months of age.

Breast milk samples were collected from the mother at the following days postpartum: 0-3 (V1), 17±3 (V2), 30±3 (V3), 60±5 days (V4), 90±5 days (V5) and 120±5 days (v6). Samples were collected after full expression from one breast using a milk pump (Symphony Breastpump, Medela), while the baby was fed on the other breast to produce a satisfactory let-down. All efforts were made to collect complete feed that included fore-, mid-, and hind-milk as a representation of one feed and to avoid within feed variation of lipid and other nutrient contents. Approximately 30 mL aliquot was separated into two conical 15 mL polypropylene tubes for analysis and the rest was returned to the mother to feed the infant. Samples collected for research were stored at −80° C. and shipped on dry ice for analyses to the Nestlé Research Center, Lausanne, Switzerland.

Information on parity (primiparous versus multiparous) were collected along with other maternal sociodemographic and anthropometric characteristics. The concentrations of HMOs were measured in breast milk at all the time points as described below.

HMO were analysed by ulta high performance liquid chromatography (UHPLC) with fluorescence detection (FLD) after labelling with anthranilamide (2AB). Milk samples (50 μL) or HMO standard solutions (50 μL) were mixed with laminaritriose solution (0.5 μmol/mL; 50 μL) used as internal standard. 2AB labelling solution (2AB, 0.35 mol/L+sodium cyanoborohydride, 1.0 mol/L in DMSO containing 30% acetic acid; 200 μL) was added and the solution heated at 65° C. for 2 h. After 2 h the samples (and standards) were cooled to 4° C. for 10 min and diluted with a solution of acetonitrile/water (75/25; 600 μL). After mixing well, the solutions were placed in a centrifuge (10000×g; 5 min) to remove particulates and the supernatant transferred to vials suitable for the UHPLC autosampler. The HMO were separated on a Waters BEH Glycan column (2.1×150 mm, 1.7 μm), preceded by a Waters BEH Amide Pre-column (2.1×5.0 mm, 1.7 μm) plumbed in to the system in such a way to act as a trapping column for removal of the excess labelling reagents (previously described by Benet & Austin, 2011) using the gradient described below. The 2AB-labelled oligosaccharides were detected by monitoring their fluorescence using λex=330 nm and λem=420 nm.

Quantification was performed against standards of the genuine HMO for 2′FL, 3FL, A-tetrasaccharide, 3′SL, 6′SL, LNT, LNnT, LNFP-I, LNFP-V, and LNnFP. All other HMO were quantified against maltotriose assuming an equimolar response of the 2AB-labelled oligosaccharides.

The following conditions were used for Separation of HMO on a BEH Glycan Column:

Time Mobile phase* 10 ports valve (min) Flow (mL/min) % A % B position Comment 0.0 0.5 95.0 5.0 10-1  Inject 5.0 μL Sample loading on trapping col. 2.3 0.5 95.0 5.0 10-1  Switch valve - start acquisition 2.5 0.5 90.0 10.0 1-2 4.9 0.5 90.0 10.0 1-2 Elution 32.1 0.5 82.0 18.0 1-2 48.1 0.5 80.5 19.5 1-2 61.5 0.5 78.0 22.0 1-2 89.0 0.5 74.6 25.4 1-2 89.5 0.4 30.0 70.0 1-2 Washing analytical col. 92.0 0.4 30.0 70.0 1-2 93.0 0.4 90.0 10.0 1-2 Re-equilibrate analytical col. 98.0 0.5 90.0 10.0 10-1  Autozero/switch valve/ stop acquisition 99.0 0.5 95.0 5.0 10-1  Equilibrate trapping col. 99.5 0.5 95.0 5.0 10-1  End

Benet, T. & Austin, S. (2011) On-line clean-up for 2-aminobenzamide-labeled oligosaccharides, Anal. Chem. 414: 166-168. http://dx.doi.org/10.1016/j.ab.2011.03.002.

The results of the compositional analysis were then subject to a statistical analysis.

A linear mixed model was used to model each HMO in which visit, parity, country and interaction between visit and parity were used as fixed effects. The within subject variability due to longitudinal repeat measures were taken care of in the model by declaring subject as a random effect.

The following statistical model was employed:

HMO^(˜)Timepoint*Parity+Country+e

Timepoint, Parity and Country refers to the fixed effects of the model and takes into consideration the interactions between timepoint and parity. e refers to the random effect of the model which controls for within subject variability.

The results of the Statistical analysis (statistical inference) are show in in tables IV-XXIV and FIGS. 1 to 19. P value tables are given in the table beneath the compound and results to which they refer.

TABLE IV param unit DELITYP visit N mean sd median q1.25% q3.75% min max 2′ - mg/L UNIQUE V1 138 4006.75 1517.06 3767.15924 3080.24 4761.75 175.61 9589.39 Fucosyllactose 2′ - mg/L UNIQUE V2 171 2715.34 976.99 2715.344 2018.57 3409.34 88.07 5848.58 Fucosyllactose 2′ - mg/L UNIQUE V3 149 2517.85 873 2442.034 1959.83 3128.59 325.18 4409.01 Fucosyllactose 2′ - mg/L UNIQUE V4 134 2122.26 813.22 2005.177885 1565.84 2658.09 189.51 4026.61 Fucosyllactose 2′ - mg/L UNIQUE V5 129 1819.24 721.8 1706.086 1335.71 2347.2 125.08 3820.31 Fucosyllactose 2′ - mg/L UNIQUE V6 122 1628.42 675.81 1526.19981 1188.35 2071.5 124.18 3608.3 Fucosyllactose 2′ - mg/L WITH V1 48 4386.09 1736.73 4289.65 3228.69 5129.99 1100.93 9498.58 Fucosyllactose SIBLINGS 2′ - mg/L WITH V2 54 2587.29 918.13 2426.2 2041.13 3043.69 533.88 5307.33 Fucosyllactose SIBLINGS 2′ - mg/L WITH V3 52 2442.11 899.71 2320.87 1799.47 2927.19 399.74 4485.03 Fucosyllactose SIBLINGS 2′ - mg/L WITH V4 50 2110.74 738.82 2022.34 1553.69 2605.41 295.32 3824.97 Fucosyllactose SIBLINGS 2′ - mg/L WITH V5 48 1894.55 712.41 1817.36 1381.61 2359.19 295.69 3621.32 Fucosyllactose SIBLINGS 2′ - mg/L WITH V6 46 1651.26 625.97 1621.59 1210.65 2096.49 159.25 2986.33 Fucosyllactose SIBLINGS

TABLE IVa Visit Estimate SE pvalue V1 382.33277 163.9708 0.01989229 V2 -134.65750 159.1302 0.39761651 V3 -141.87577 161.5152 0.37991228 V4 -152.21420 163.7197 0.35271384 V5 -90.77581 165.3151 0.58312380 V6 -169.04976 166.9656 0.31152596

TABLE V param unit DELITYP visit N mean sd median q1.25% q3.75% min max 3 - mg/L UNIQUE V1 172 385.4664991 415.5565524 237.86708 148.2090175 398.264 11.41228 2468.566 Fucosyl- lactose 3 - mg/L UNIQUE V2 214 553.0388569 521.3096535 354.48191 210.854 649.0720075 43.78025 2537.227 Fucosyl- lactose 3 - mg/L UNIQUE V3 189 674.7012092 574.5232909 478.311 275.97259 831.444 57.45987 2921.70036 Fucosyl- lactose 3 - mg/L UNIQUE V4 173 928.8239356 668.6906753 708.247 454.831 1209.657 81.083 3278.66994 Fucosyl- lactose 3 - mg/L UNIQUE V5 166 1086.633496 757.0840296 864.07916 571.399 1410.950658 93.05087 5715.55268 Fucosyl- lactose 3 - mg/L UNIQUE V6 158 1160.839661 682.5011962 998.615 621.92175 1606.707478 112.962 3443.91 Fucosyl- lactose 3 - mg/L WITH V1 64 519.4932994 531.8511225 289.8366 169.507215 728.1158025 32.664 2078.125 Fucosyl- SIBLINGS lactose 3 - mg/L WITH V2 75 711.2907307 628.9001769 441.16265 299.365935 939.567185 40.313 2638.36113 Fucosyl- SIBLINGS lactose 3 - mg/L WITH V3 71 839.9917762 678.0150354 633.44927 345.148815 1173.1234 58.9511 2670.76 Fucosyl- SIBLINGS lactose 3 - mg/L WITH V4 69 1073.123048 742.4445881 937.19072 548.39844 1322.12 78.17999 2963.532 Fucosyl- SIBLINGS lactose 3 - mg/L WITH V5 67 1271.12714 815.3119729 1038.34043 672.70648 1623.508555 84.97374 3499.62987 Fucosyl- SIBLINGS lactose 3 - mg/L WITH V6 65 1327.515428 787.8852486 1196.455 757.36606 1674.172 109.34912 3653.15978 Fucosyl- SIBLINGS lactose

TABLE VI Visit Estimate SE pvalue V1 1.304577 1.116104 0.01561971 V2 1.270724 1.114634 0.02742312 V3 1.249569 1.115363 0.04146459 V4 1.189985 1.1157696 0.11261200 V5 1.180027 1.116120 0.13207376 V6 1.169900 1.116435 0.15445449

TABLE VII param unit DELITYP visit N mean sd median q1.25% 6′ - mg/L UNIQUE V1 173 128.9940719 47.46982 126.3577251 100.3019356 Galactosyllactose 6′ - mg/L UNIQUE V2 214 40.43950928 34.32453 34.93363996 27.84100032 Galactosyllactose 6′ - mg/L UNIQUE V3 190 25.44688677 9.806509 23.17565201 19.42520304 Galactosyllactose 6′ - mg/L UNIQUE V4 173 17.59295944 8.05038 15.74108242 12.47470354 Galactosyllactose 6′ - mg/L UNIQUE V5 161 14.34098883 7.479945 12.4716807 10.16207027 Galactosyllactose 6′ - mg/L UNIQUE V6 147 13.07171018 12.49064 10.80183408 8.87243724 Galactosyllactose 6′ - mg/L WITH V1 64 138.9473035 46.10431 137.0616097 106.2274385 Galactosyllactose SIBLINGS 6′ - mg/L WITH V2 74 39.1280561 15.2604 35.89756199 29.57195798 Galactosyllactose SIBLINGS 6′- mg/L WITH V3 71 26.40557425 10.78379 24.87935762 20.45856916 Galactosyllactose SIBLINGS 6′ - mg/L WITH V4 67 18.19156281 9.872754 16.27389683 12.55895567 Galactosyllactose SIBLINGS 6′ - mg/L WITH V5 65 14.43524793 7.081825 12.01385773 9.826798342 Galactosyllactose SIBLINGS 6′ - mg/L WITH V6 61 12.24780197 5.723957 10.93564962 9.250831953 Galactosyllactose SIBLINGS param unit DELITYP visit q3.75% min max 6′ - mg/L UNIQUE V1 154.0188447 11.28896522 288.7462236 Galactosyllactose 6′ - mg/L UNIQUE V2 42.73797305 13.62127529 434.3117534 Galactosyllactose 6′ - mg/L UNIQUE V3 29.16381402 7.291378016 88.87964314 Galactosyllactose 6′ - mg/L UNIQUE V4 19.96432738 6.838216821 65.30491452 Galactosyllactose 6′ - mg/L UNIQUE V5 16.09249333 6.514312709 60.46113641 Galactosyllactose 6′ - mg/L UNIQUE V6 13.69842265 6.482990651 146.8862427 Galactosyllactose 6′ - mg/L WITH V1 168.6023267 29.69176747 258.227941 Galactosyllactose SIBLINGS 6′ - mg/L WITH V2 44.46636476 14.16036621 98.52362497 Galactosyllactose SIBLINGS 6′- mg/L WITH V3 29.31283111 12.8653756 85.11500318 Galactosyllactose SIBLINGS 6′ - mg/L WITH V4 21.00367988 6.668169255 74.29089736 Galactosyllactose SIBLINGS 6′ - mg/L WITH V5 17.61023478 7.170486535 55.96674008 Galactosyllactose SIBLINGS 6′ - mg/L WITH V6 13.5930005 6.800981073 43.62702667 Galactosyllactose SIBLINGS

TABLE VIII param unit DELITYP visit N mean sd median q1.25% q3.75% min max BloodgroupA- mg/L UNIQUE V1 62 218.3100965 199.91 137.645 96.501955 339.8993275 16.70496 1198.809 Tetrasaccharide BloodgroupA- mg/L UNIQUE V2 74 108.9752193 87.04024 73.80618 49.86775 153.6967025 20.571 406.27274 Tetrasaccharide BloodgroupA- mg/L UNIQUE V3 67 101.1101424 80.23 74.20433 44.016415 113.37141 18.04427 388.594 Tetrasaccharide BloodgroupA- mg/L UNIQUE V4 59 89.30594305 70.23909 58.313 39.61766 128.653605 15.067 338.339 Tetrasaccharide BloodgroupA- mg/L UNIQUE V5 53 85.24264679 64.13099 62.96622 35.832 119.28 17.93698 269.711 Tetrasaccharide BloodgroupA- mg/L UNIQUE V6 48 78.27483 59.40327 54.919015 32.0109175 113.239025 15.08493 214.554 Tetrasaccharide BloodgroupA- mg/L WITH V1 18 224.7452844 294.2047 120.82463 35.8747 284.1568725 16.98872 993.8064 Tetrasaccharide SIBLINGS BloodgroupA- mg/L WITH V2 18 112.8672322 117.5963 75.509785 48.8410875 159.6912825 15.208 517.54089 Tetrasaccharide SIBLINGS BloodgroupA- mg/L WITH V3 17 123.0597341 119.73 95.024 48.21369 146.4399 17.56554 529.36 Tetrasaccharide SIBLINGS BloodgroupA- mg/L WITH V4 17 98.30965941 83.89327 77.95666 57.209 115.87755 16.361 376.29878 Tetrasaccharide SIBLINGS BloodgroupA- mg/L WITH V5 15 85.03220533 69.04539 67.01464 40.436035 107.519645 19.39048 294.134 Tetrasaccharide SIBLINGS BloodgroupA- mg/L WITH V6 15 70.211992 56.09551 51.54932 35.436115 88.07794 21.94559 246.20116 Tetrasaccharide SIBLINGS

TABLE IX param unit DELITYP visit N mean sd median q1.25% Difucosyllacto-N- mg/L UNIQUE V1 125 166.66388 100.4515893 150.4463936 98.1823431 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V2 167 282.797729 161.029439 247.1195693 178.1422564 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V3 144 234.3063825 147.4320416 200.969341 134.8599339 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V4 118 119.7067972 96.5242152 94.42787799 52.25253247 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V5 78 98.51640533 80.74105435 72.88240189 50.34510842 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V6 58 74.99587394 51.32117734 59.0456174 42.88833806 Hexaose-a Difucosyllacto-N- mg/L WITH V1 48 148.346054 81.70434542 125.6285912 94.72884661 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V2 54 261.9050265 169.8874876 231.2689262 167.1776416 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V3 51 206.2678072 144.2391624 170.0672888 110.5643357 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V4 42 120.121861 100.4379777 102.9007162 56.84164741 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V5 29 97.3857747 77.50806012 71.724665 59.86775755 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V6 24 76.45717107 53.3864606 65.56011998 42.44962707 Hexaose-a SIBLINGS param unit DELITYP visit q3.75% min max Difucosyllacto-N- mg/L UNIQUE V1 208.71468 37.68514348 576.8930576 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V2 355.9677629 33.08461539 856.5390082 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V3 300.7426145 39.43414695 807.2575598 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V4 149.7452022 33.55783812 538.5644379 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V5 107.3617675 33.27717717 515.2208712 Hexaose-a Difucosyllacto-N- mg/L UNIQUE V6 82.61864327 33.45887998 270.1883637 Hexaose-a Difucosyllacto-N- mg/L WITH V1 188.3725375 33.04833329 412.2467197 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V2 307.2992152 41.111123 1086.897955 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V3 253.965274 42.14158522 747.7649157 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V4 127.1671806 34.70128067 606.8605662 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V5 89.40771747 33.00609983 393.74294 Hexaose-a SIBLINGS Difucosyllacto-N- mg/L WITH V6 80.98116787 33.43153153 266.5264593 Hexaose-a SIBLINGS

TABLE X param unit DELITYP visit N Mean sd median Lacto-N- mg/L UNIQUE V1 126 1234.406373 535.3466 1180.198793 Difucosylhexaose Lacto-N- mg/L UNIQUE V2 158 1255.755653 537.4429 1192.294603 Difucosylhexaose Lacto-N- mg/L UNIQUE V3 136 1091.823028 465.8642 995.3117755 Difucosylhexaose Lacto-N- mg/L UNIQUE V4 121 848.0091077 328.5295 802.3504693 Difucosylhexaose Lacto-N- mg/L UNIQUE V5 116 710.6302809 271.9261 681.6194012 Difucosylhexaose Lacto-N- mg/L UNIQUE V6 112 634.9542185 242.0775 624.6350378 Difucosylhexaose Lacto-N- mg/L WITH V1 46 1225.800254 477.4247 1169.458449 Difucosylhexaose SIBLINGS Lacto-N- mg/L WITH V2 51 1336.18374 578.9028 1348.485217 Difucosylhexaose SIBLINGS Lacto-N- mg/L WITH V3 48 1142.006226 411.3581 1114.291686 Difucosylhexaose SIBLINGS Lacto-N- mg/L WITH V4 48 825.7560502 324.6839 788.8361782 Difucosylhexaose SIBLINGS Lacto-N- mg/L WITH V5 45 739.1373379 317.1183 701.7743271 Difucosylhexaose SIBLINGS Lacto-N- mg/L WITH V6 44 578.2746578 236.0452 567.1716751 Difucosylhexaose SIBLINGS param q1.25% q3.75% min max Lacto-N- 849.3209761 1495.918413 11.58325101 3346.954498 Difucosylhexaose Lacto-N- 942.9128926 1550.664139 21.63261134 3582.842867 Difucosylhexaose Lacto-N- 821.2656843 1344.286596 10.97928599 3501.352786 Difucosylhexaose Lacto-N- 662.7755195 1023.609883 11.88700135 1649.120744 Difucosylhexaose Lacto-N- 541.1164043 837.2962088 103.063301 1722.213919 Difucosylhexaose Lacto-N- 471.4977544 756.5617445 111.6094256 1485.00214 Difucosylhexaose Lacto-N- 878.4507667 1407.01817 462.8830063 2782.944079 Difucosylhexaose Lacto-N- 1108.263831 1576.688066 12.29676207 3031.332599 Difucosylhexaose Lacto-N- 884.4304615 1431.628644 169.4986155 1927.975206 Difucosylhexaose Lacto-N- 631.0881501 1035.617596 74.64280102 1917.735932 Difucosylhexaose Lacto-N- 536.5070919 882.8586901 221.5440208 1793.170075 Difucosylhexaose Lacto-N- 413.7894666 739.2552491 60.80285161 1261.115824 Difucosylhexaose

TABLE XI param unit DELITYP visit N Mean sd median Lacto-N- mg/L UNIQUE V1 139 1953.405264 895.9388 1913.01447 Fucosylpentaose-I Lacto-N- mg/L UNIQUE V2 170 1442.925271 803.4876 1369.9995 Fucosylpentaose-I Lacto-N- mg/L UNIQUE V3 149 1080.473899 640.1305 933.32871 Fucosylpentaose-I Lacto-N- mg/L UNIQUE V4 133 610.916764 413.7517 503.48892 Fucosylpentaose-I Lacto-N- mg/L UNIQUE V5 127 470.6971887 378.858 376.267 Fucosylpentaose-I Lacto-N- mg/L UNIQUE V6 120 388.8400843 320.5121 315.61007 Fucosylpentaose-I Lacto-N- mg/L WITH V1 48 1855.27654 929.5294 1704.10064 Fucosylpentaose-I SIBLINGS Lacto-N- mg/L WITH V2 54 1391.449066 788.47 1172.082955 Fucosylpentaose-I SIBLINGS Lacto-N- mg/L WITH V3 52 1044.350567 593.307 1006.408885 Fucosylpentaose-I SIBLINGS Lacto-N- mg/L WITH V4 50 611.3230378 451.9114 562.65468 Fucosylpentaose-I SIBLINGS Lacto-N- mg/L WITH V5 48 465.4264017 359.7624 380.606735 Fucosylpentaose-I SIBLINGS Lacto-N- mg/L WITH V6 45 370.5976704 304.8824 293.10456 Fucosylpentaose-I SIBLINGS param q1.25% q3.75% min max Lacto-N- 1330.993545 2520.18355 27.4573 4040.89442 Fucosylpentaose-I Lacto-N- 781.2125 2003.57863 49.806 3756.06871 Fucosylpentaose-I Lacto-N- 573.47716 1496.567 28.866 3306.848 Fucosylpentaose-I Lacto-N- 283.029 832.54689 30.669 1970.645 Fucosylpentaose-I Lacto-N- 185.98 617.2625 30.394 2062.94583 Fucosylpentaose-I Lacto-N- 161.52072 501.8237575 27.51296 1913.34495 Fucosylpentaose-I Lacto-N- 1202.36626 2417.197 304.92323 4311.24617 Fucosylpentaose-I Lacto-N- 808.9399325 1909.67225 106.64425 3571.06665 Fucosylpentaose-I Lacto-N- 594.47375 1446.30243 77.48494 2372.66273 Fucosylpentaose-I Lacto-N- 285.0165 781.02097 36.497 2068.53637 Fucosylpentaose-I Lacto-N- 194.196455 608.467225 28.33995 1643.26 Fucosylpentaose-I Lacto-N- 153.08 490.82 43.879 1622.958 Fucosylpentaose-I

TABLE XII param unit DELITYP visit N mean sd median Lacto-N- mg/L UNIQUE V1 145 400.7219326 518.1076 192.6370398 Fucosylpentaose-II Lacto-N- mg/L UNIQUE V2 190 544.0406658 590.6689 327.9102556 Fucosylpentaose-II Lacto-N- mg/L UNIQUE V3 165 512.6256601 521.4556 320.0085042 Fucosylpentaose-II Lacto-N- mg/L UNIQUE V4 149 461.1191582 396.6782 331.221059 Fucosylpentaose-II Lacto-N- mg/L UNIQUE V5 146 420.9635001 326.0906 332.4659875 Fucosylpentaose-II Lacto-N- mg/L UNIQUE V6 138 383.5239207 276.9464 293.9566112 Fucosylpentaose-II Lacto-N- mg/L WITH V1 57 474.9349989 519.0685 248.5627023 Fucosylpentaose-II SIBLINGS Lacto-N- mg/L WITH V2 70 732.4415102 712.1251 428.9741842 Fucosylpentaose-II SIBLINGS Lacto-N- mg/L WITH V3 65 642.6995951 550.2848 458.7201961 Fucosylpentaose-II SIBLINGS Lacto-N- mg/L WITH V4 64 504.6233294 416.96 327.235739 Fucosylpentaose-II SIBLINGS Lacto-N- mg/L WITH V5 63 461.6140679 346.8066 347.3093904 Fucosylpentaose-II SIBLINGS Lacto-N- mg/L WITH V6 61 416.7272357 300.1685 326.0899963 Fucosylpentaose-II SIBLINGS param q1.25% q3.75% min max Lacto-N- 103.6228597 323.2837721 37.04755787 2426.895823 Fucosylpentaose-II Lacto-N- 182.5788937 568.883943 52.61106978 2856.419409 Fucosylpentaose-II Lacto-N- 184.3943767 543.5270238 53.24693544 2488.674427 Fucosylpentaose-II Lacto-N- 216.7351314 562.6200229 60.89431943 2274.458362 Fucosylpentaose-II Lacto-N- 200.0765447 498.1834633 53.63244898 1714.785797 Fucosylpentaose-II Lacto-N- 215.2587313 449.8741781 50.47498562 1554.445901 Fucosylpentaose-II Lacto-N- 124.7083137 752.9452298 38.05551582 2146.380077 Fucosylpentaose-II Lacto-N- 256.8757435 1123.163255 35.31341987 3602.345161 Fucosylpentaose-II Lacto-N- 240.4601421 955.8715703 88.28875496 2623.432553 Fucosylpentaose-II Lacto-N- 217.4510082 639.6884927 35.93501895 2091.202327 Fucosylpentaose-II Lacto-N- 231.9723537 592.70929 41.55891583 1551.01748 Fucosylpentaose-II Lacto-N- 214.4554786 547.9185395 44.29386143 1488.541991 Fucosylpentaose-II

TABLE XIII Visit Estimate SE pvalue V1 1.276220 1.132216 0.04972522 V2 1.350333 1.130009 0.01412686 V3 1.337439 1.131037 0.01835975 V4 1.211846 1.131458 0.12001568 V5 1.191473 1.131706 0.15703207 V6 1.147623 1.132101 0.26730874

TABLE XIV param unit DELITYP visit N mean sd median Lacto-N- mg/L UNIQUE V1 172 440.7436567 162.9639 425.9454976 Fucosylpentaose-III Lacto-N- mg/L UNIQUE V2 214 324.5042397 155.5033 307.6437027 Fucosylpentaose-III Lacto-N- mg/L UNIQUE V3 189 313.7021468 104.9384 304.1777874 Fucosylpentaose-III Lacto-N- mg/L UNIQUE V4 173 361.8867399 117.8097 352.9061564 Fucosylpentaose-III Lacto-N- mg/L UNIQUE V5 166 355.1420518 92.17895 351.5526117 Fucosylpentaose-III Lacto-N- mg/L UNIQUE V6 158 345.2252395 93.83141 337.6224468 Fucosylpentaose-III Lacto-N- mg/L WITH V1 64 454.7211229 173.722 441.4279491 Fucosylpentaose-III SIBLINGS Lacto-N- mg/L WITH V2 75 307.1033885 86.78954 304.261907 Fucosylpentaose-III SIBLINGS Lacto-N- mg/L WITH V3 71 303.1843203 78.66454 293 Fucosylpentaose-III SIBLINGS Lacto-N- mg/L WITH V4 69 348.8290902 89.00072 335.6039586 Fucosylpentaose-III SIBLINGS Lacto-N- mg/L WITH V5 67 349.2434082 93.08592 345.6968971 Fucosylpentaose-III SIBLINGS Lacto-N- mg/L WITH V6 65 323.8386581 87.5295 313.2473169 Fucosylpentaose-III SIBLINGS param q1.25% q3.75% min max Lacto-N- 326.9628144 523.1014552 117.4373288 1151.993123 Fucosylpentaose-III Lacto-N- 231.1082286 388.025127 79.94840168 1751.020203 Fucosylpentaose-III Lacto-N- 252.945844 378.3723403 97.1179054 951.8773833 Fucosylpentaose-III Lacto-N- 289.4064649 414.3069494 80.61587114 1196.494936 Fucosylpentaose-III Lacto-N- 281.5673515 416.9454623 166.2976602 635.2849534 Fucosylpentaose-III Lacto-N- 284.583471 413.5865187 138.1858699 557.1545027 Fucosylpentaose-III Lacto-N- 315.3262438 541.2156412 187.9371289 890.3129598 Fucosylpentaose-III Lacto-N- 246.1138864 356.6258239 87.25376406 535.9760196 Fucosylpentaose-III Lacto-N- 257.845359 348.19862 147.4060444 563.392227 Fucosylpentaose-III Lacto-N- 292.1571655 405.4241552 160.7903865 543.5231017 Fucosylpentaose-III Lacto-N- 278.5226687 399.4384528 126.2259899 679.9790614 Fucosylpentaose-III Lacto-N- 251.2469262 383.3281509 134.2613528 567.5952348 Fucosylpentaose-III

TABLE XV param unit DELITYP visit N mean sd median Lacto-N- mg/L UNIQUE V1 112 109.4842732 107.2949 48.9985 Fucosylpentaose-V Lacto-N- mg/L UNIQUE V2 175 119.017012 114.1155 63.592 Fucosylpentaose-V Lacto-N- mg/L UNIQUE V3 155 109.9870335 101.3161 61.36453 Fucosylpentaose-V Lacto-N- mg/L UNIQUE V4 144 90.07381556 72.94248 58.491385 Fucosylpentaose-V Lacto-N- mg/L UNIQUE V5 135 87.41945786 70.56107 57.784 Fucosylpentaose-V Lacto-N- mg/L UNIQUE V6 129 76.81346488 59.9275 54.037 Fucosylpentaose-V Lacto-N- mg/L WITH V1 46 105.4405772 91.79857 57.939865 Fucosylpentaose-V SIBLINGS Lacto-N- mg/L WITH V2 68 138.2168362 125.1875 84.83034 Fucosylpentaose-V SIBLINGS Lacto-N- mg/L WITH V3 65 116.1223805 95.20984 75.977 Fucosylpentaose-V SIBLINGS Lacto-N- mg/L WITH V4 62 91.59901602 71.28318 64.315775 Fucosylpentaose-V SIBLINGS Lacto-N- mg/L WITH V5 60 79.1156255 55.1115 59.570785 Fucosylpentaose-V SIBLINGS Lacto-N- mg/L WITH V6 57 73.00692544 49.32591 53.91017 Fucosylpentaose-V SIBLINGS param q1.25% q3.75% min max Lacto-N- 34.0883725 176.98825 24.098 393.23939 Fucosylpentaose-V Lacto-N- 42.54693 172.5235 24.15465 495.7132 Fucosylpentaose-V Lacto-N- 44.732165 132.402505 24.357 427.275 Fucosylpentaose-V Lacto-N- 41.540815 120.62295 24.31422 412.37003 Fucosylpentaose-V Lacto-N- 42.07251 105.3295 25.775 360.11687 Fucosylpentaose-V Lacto-N- 39.65592 91.13553 24.172 325 Fucosylpentaose-V Lacto-N- 36.8393575 165.8721925 24.184 309.94073 Fucosylpentaose-V Lacto-N- 49.35551 224.66925 25.57568 515.4305 Fucosylpentaose-V Lacto-N- 50.50503 148.75439 24.72194 414.4 Fucosylpentaose-V Lacto-N- 41.231875 111.3105 24.48 360.47113 Fucosylpentaose-V Lacto-N- 38.57574 101.87361 26.214 243.549 Fucosylpentaose-V Lacto-N- 38.03945 97.496 24.586 232.777 Fucosylpentaose-V

TABLE XVI param unit DELITYP visit N Mean Sd median Lacto-N-hexaose mg/L UNIQUE V1 139 69.65726947 60.09699 58.2952622 (A) Lacto-N-hexaose mg/L UNIQUE V2 205 83.91299646 44.09424 72.90325819 (A) Lacto-N-hexaose mg/L UNIQUE V3 185 71.67837167 65.1411 60.20319698 (A) Lacto-N-hexaose mg/L UNIQUE V4 154 39.62268882 18.54261 34.7920641 (A) Lacto-N-hexaose mg/L UNIQUE V5 130 30.26720457 14.06454 25.06469316 (A) Lacto-N-hexaose mg/L UNIQUE V6 107 25.29538373 9.751377 21.98491257 (A) Lacto-N-hexaose mg/L WITH V1 53 69.44083736 43.31972 62.07164331 (A) SIBLINGS Lacto-N-hexaose mg/L WITH V2 74 69.18762737 31.78699 60.56720348 (A) SIBLINGS Lacto-N-hexaose mg/L WITH V3 70 62.49890449 31.54116 53.23506574 (A) SIBLINGS Lacto-N-hexaose mg/L WITH V4 63 38.80460416 20.67884 34.70663487 (A) SIBLINGS Lacto-N-hexaose mg/L WITH V5 52 33.10356562 20.12426 27.47100266 (A) SIBLINGS Lacto-N-hexaose mg/L WITH V6 51 27.65797971 11.52966 24.0510901 (A) SIBLINGS param q1.25% q3.75% min max Lacto-N-hexaose 38.17826996 81.61276425 18.36222878 585.5330588 (A) Lacto-N-hexaose 54.70601626 109.8055481 17.46965778 255.4484418 (A) Lacto-N-hexaose 43.02753693 84.88738297 16.19895295 803.0893219 (A) Lacto-N-hexaose 27.13590373 46.62997118 16.3928703 112.8544379 (A) Lacto-N-hexaose 20.88531751 32.23440104 16.0313543 87.57130429 (A) Lacto-N-hexaose 19.09685889 27.93809463 16.1076984 68.12521273 (A) Lacto-N-hexaose 46.00691942 81.83799145 16.96457388 248.8007182 (A) Lacto-N-hexaose 45.91588703 82.53308105 18.15175655 169.5397907 (A) Lacto-N-hexaose 41.84623544 77.85927254 17.24348703 202.4682182 (A) Lacto-N-hexaose 26.44253846 44.29626051 17.87155202 139.2964627 (A) Lacto-N-hexaose 22.26315853 38.02796933 16.26681559 141.4095103 (A) Lacto-N-hexaose 20.30278179 31.43146137 16.26239728 69.64415593 (A)

TABLE XVII param unit DELITYP Visit N mean Sd median Lacto-N-hexaose mg/L UNIQUE V1 134 44.73295024 34.32684 33.28233637 (B) Lacto-N-hexaose mg/L UNIQUE V2 145 43.11011358 23.31967 35.13295515 (B) Lacto-N-hexaose mg/L UNIQUE V3 133 38.50327162 21.12716 32.35290783 (B) Lacto-N-hexaose mg/L UNIQUE V4 91 33.26540708 18.18317 26.42650386 (B) Lacto-N-hexaose mg/L UNIQUE V5 59 31.002883 17.26554 24.99800181 (B) Lacto-N-hexaose mg/L UNIQUE V6 43 27.72187917 15.68486 22.97054189 (B) Lacto-N-hexaose mg/L WITH V1 40 41.5669005 25.93896 37.79728256 (B) SIBLINGS Lacto-N-hexaose mg/L WITH V2 36 36.02515556 16.13595 30.58969053 (B) SIBLINGS Lacto-N-hexaose mg/L WITH V3 43 34.28955775 15.68827 29.61018115 (B) SIBLINGS Lacto-N-hexaose mg/L WITH V4 29 34.74970859 13.55697 31.13853753 (B) SIBLINGS Lacto-N-hexaose mg/L WITH V5 22 28.51987306 13.09366 24.76177376 (B) SIBLINGS Lacto-N-hexaose mg/L WITH V6 18 28.88319141 11.91772 27.2941175 (B) SIBLINGS param q1.25% q3.75% min max Lacto-N-hexaose 22.79344461 53.48647374 16.03905983 220.7927384 (B) Lacto-N-hexaose 27.90776152 55.01012587 16.31270557 179.8286126 (B) Lacto-N-hexaose 24.09883189 45.67418316 16.18714448 131.3072962 (B) Lacto-N-hexaose 19.69922881 39.64344687 16.41060296 115.2056418 (B) Lacto-N-hexaose 20.04178856 33.98491795 16.25904523 105.2379451 (B) Lacto-N-hexaose 18.6143141 32.997549 16.02806725 99.84562336 (B) Lacto-N-hexaose 24.8190705 44.77882074 16.60528968 135.9994959 (B) Lacto-N-hexaose 23.74009313 46.45568574 17.15234171 84.77670516 (B) Lacto-N-hexaose 23.11258219 45.36245075 16.5380127 85.67908288 (B) Lacto-N-hexaose 24.25250318 43.6851328 16.78734358 76.04017383 (B) Lacto-N-hexaose 18.2128352 33.18017398 16.04481676 63.3735739 (B) Lacto-N-hexaose 20.37699792 31.56511312 16.83515397 62.67504542 (B)

TABLE XVIII param unit DELITYP Visit N mean sd median Lacto-N- mg/L UNIQUE V1 117 111.9565166 69.43106 94.19356953 Neodifucosylhexaose Lacto-N- mg/L UNIQUE V2 69 63.29788709 74.07394 43.4578365 Neodifucosylhexaose Lacto-N- mg/L UNIQUE V3 43 57.46798821 56.84744 46.51065959 Neodifucosylhexaose Lacto-N- mg/L UNIQUE V4 56 55.60053979 33.48608 41.85130914 Neodifucosylhexaose Lacto-N- mg/L UNIQUE V5 38 54.58027932 27.91287 45.11014515 Neodifucosylhexaose Lacto-N- mg/L UNIQUE V6 29 64.33423068 33.2452 56.79116124 Neodifucosylhexaose Lacto-N- mg/L WITH V1 46 114.8602548 82.49724 80.63002523 Neodifucosylhexaose SIBLINGS Lacto-N- mg/L WITH V2 19 61.94019548 45.97203 45.4381543 Neodifucosylhexaose SIBLINGS Lacto-N- mg/L WITH V3 15 58.61383439 39.13784 54.97433839 Neodifucosylhexaose SIBLINGS Lacto-N- mg/L WITH V4 19 54.94306309 22.66628 48.37428768 Neodifucosylhexaose SIBLINGS Lacto-N- mg/L WITH V5 18 63.29320064 32.14269 53.26006424 Neodifucosylhexaose SIBLINGS Lacto-N- mg/L WITH V6 16 58.9507447 27.15766 45.11453324 Neodifucosylhexaose SIBLINGS param q1.25% q3.75% min max Lacto-N- 59.70978875 132.6595225 29.32389989 334.1268818 Neodifucosylhexaose Lacto-N- 36.27672844 68.9398609 28.52752291 612.4524307 Neodifucosylhexaose Lacto-N- 32.51100498 65.227353 28.13123604 398.4920634 Neodifucosylhexaose Lacto-N- 34.6297264 73.17992495 28.10296364 224.0399443 Neodifucosylhexaose Lacto-N- 34.29570779 63.77490019 28.12614506 141.5694861 Neodifucosylhexaose Lacto-N- 35.66139704 82.82903182 28.64848952 151.8818948 Neodifucosylhexaose Lacto-N- 57.15833441 153.8645579 31.26569753 350.3302321 Neodifucosylhexaose Lacto-N- 35.75460343 68.64819832 29.95124997 226.4465075 Neodifucosylhexaose Lacto-N- 34.52825063 66.41893776 29.35106887 187.0453095 Neodifucosylhexaose Lacto-N- 40.02309424 65.92514047 28.49364998 114.5744313 Neodifucosylhexaose Lacto-N- 38.88978245 79.91072693 30.09514734 140.8977209 Neodifucosylhexaose Lacto-N- 39.24229995 73.74432573 32.02848488 111.8547311 Neodifucosylhexaose

TABLE XIX param unit DELITYP visit N mean sd median Lacto-N- mg/L UNIQUE V1 50 37.2525136 16.38068 33.67124 Neofucosylpentaose Lacto-N- mg/L UNIQUE V2 42 29.88595143 10.58937 26.92816 Neofucosylpentaose Lacto-N- mg/L UNIQUE V3 40 27.0749745 9.243692 23.80713 Neofucosylpentaose Lacto-N- mg/L UNIQUE V4 39 30.60720744 12.18325 27.47212 Neofucosylpentaose Lacto-N- mg/L UNIQUE V5 31 27.02506516 6.34542 25.69 Neofucosylpentaose Lacto-N- mg/L UNIQUE V6 23 26.7532187 7.552031 24.091 Neofucosylpentaose Lacto-N- mg/L WITH V1 22 35.40536864 16.21748 28.809265 Neofucosylpentaose SIBLINGS Lacto-N- mg/L WITH V2 16 23.29734 3.547994 23.303695 Neofucosylpentaose SIBLINGS Lacto-N- mg/L WITH V3 11 31.39661 8.042168 29.12481 Neofucosylpentaose SIBLINGS Lacto-N- mg/L WITH V4 12 32.5542 11.27676 32.015775 Neofucosylpentaose SIBLINGS Lacto-N- mg/L WITH V5 11 28.90526364 10.61775 25.04186 Neofucosylpentaose SIBLINGS Lacto-N- mg/L WITH V6 9 30.85879 15.08243 24.315 Neofucosylpentaose SIBLINGS param q1.25% q3.75% min max Lacto-N- 24.897 41.76092 19.73896 92.18025 Neofucosylpentaose Lacto-N- 22.144305 30.94125 19.066 59.578 Neofucosylpentaose Lacto-N- 21.4349075 28.11675 19.002 53.52264 Neofucosylpentaose Lacto-N- 23.1454 32.614335 19.074 82.885 Neofucosylpentaose Lacto-N- 22.51126 28.6975 19.037 42.33404 Neofucosylpentaose Lacto-N- 20.97355 29.805 19.054 48.112 Neofucosylpentaose Lacto-N- 26.059 39.3506325 20.14561 84.56933 Neofucosylpentaose Lacto-N- 20.0572975 25.788 19.148 30.76332 Neofucosylpentaose Lacto-N- 25.681345 36.948515 22.29 46.081 Neofucosylpentaose Lacto-N- 23.654625 35.353345 22.437 60.886 Neofucosylpentaose Lacto-N- 21.985635 31.76967 20.88696 57.46976 Neofucosylpentaose Lacto-N- 20.677 29.41272 20.15457 64.87128 Neofucosylpentaose

TABLE XX param unit DELITYP visit N mean sd median Lacto-N- mg/L UNIQUE V1 173 307.7437813 131.4372 293.564 Neotetraose Lacto-N- mg/L UNIQUE V2 213 184.6575148 100.9842 173.50436 Neotetraose Lacto-N- mg/L UNIQUE V3 187 156.7024487 82.56116 146.70853 Neotetraose Lacto-N- mg/L UNIQUE V4 168 128.4345314 83.21485 107.724735 Neotetraose Lacto-N- mg/L UNIQUE V5 158 107.643116 69.56969 91.5955 Neotetraose Lacto-N- mg/L UNIQUE V6 146 98.41257377 60.79302 83.588435 Neotetraose Lacto-N- mg/L WITH V1 64 304.2184716 135.7726 298.56318 Neotetraose SIBLINGS Lacto-N- mg/L WITH V2 72 154.2337461 79.88463 141.929085 Neotetraose SIBLINGS Lacto-N- mg/L WITH V3 68 144.1930094 71.03133 132.30853 Neotetraose SIBLINGS Lacto-N- mg/L WITH V4 64 126.1508667 72.02305 112.7993 Neotetraose SIBLINGS Lacto-N- mg/L WITH V5 60 107.7504913 60.55004 93.494 Neotetraose SIBLINGS Lacto-N- mg/L WITH V6 62 96.30362226 64.02489 79.572765 Neotetraose SIBLINGS param q1.25% q3.75% min max Lacto-N- 214.326 389.59097 47.75439 699.27383 Neotetraose Lacto-N- 102.93676 246.335 25.73309 597.13727 Neotetraose Lacto-N- 89.75674 205.208 26.64085 424.58858 Neotetraose Lacto-N- 69.01625 167.72524 25.0473 463.904 Neotetraose Lacto-N- 58.5215 136.18827 25.20536 398.9075 Neotetraose Lacto-N- 52.0978825 121.5750225 25.22179 298.346 Neotetraose Lacto-N- 219.730275 373.411605 64.40836 695.668 Neotetraose Lacto-N- 96.1692475 202.7789125 32.626 407.85 Neotetraose Lacto-N- 85.0371325 200.53189 31.748 333.83 Neotetraose Lacto-N- 63.8390525 179.90584 25.575 309.45348 Neotetraose Lacto-N- 62.7070925 147.2496125 31.655 257.58145 Neotetraose Lacto-N- 47.7260275 118.86836 24.222 277.71054 Neotetraose

TABLE XXI param unit DELITYP visit N mean Sd median Lacto-N-Tetraose mg/L UNIQUE V1 173 941.3105777 869.2799 668.05 Lacto-N-Tetraose mg/L UNIQUE V2 215 1198.301294 760.0552 1066.41302 Lacto-N-Tetraose mg/L UNIQUE V3 190 1013.002805 642.5475 860.403755 Lacto-N-Tetraose mg/L UNIQUE V4 174 710.5043052 451.1813 624.944005 Lacto-N-Tetraose mg/L UNIQUE V5 167 617.336245 439.7945 499.74 Lacto-N-Tetraose mg/L UNIQUE V6 159 541.9378956 396.6598 460.991 Lacto-N-Tetraose mg/L WITH V1 64 833.6287748 579.2782 791.530885 SIBLINGS Lacto-N-Tetraose mg/L WITH V2 75 1255.222192 591.7982 1121.872 SIBLINGS Lacto-N-Tetraose mg/L WITH V3 71 1000.002187 425.3526 947.02993 SIBLINGS Lacto-N-Tetraose mg/L WITH V4 68 672.2036891 307.2625 647.973965 SIBLINGS Lacto-N-Tetraose mg/L WITH V5 67 552.031873 275.787 479.85205 SIBLINGS Lacto-N-Tetraose mg/L WITH V6 63 486.68108 230.7009 413.22725 SIBLINGS param q1.25% q3.75% min max Lacto-N-Tetraose 389.089 1247.059 21.9529 6714.31284 Lacto-N-Tetraose 672.8633 1443.05473 125.055 5360.81507 Lacto-N-Tetraose 608.09198 1259.128325 119.024 4011.92765 Lacto-N-Tetraose 429.9219475 815.3775375 121.34189 2894.36631 Lacto-N-Tetraose 349.65877 738.019535 85.62092 2932.955 Lacto-N-Tetraose 300.9595 640.624545 101.62557 3026.125 Lacto-N-Tetraose 402.6631225 1063.8035 53.0063 2461.67215 Lacto-N-Tetraose 840.66133 1577.087795 161.241 3144.60141 Lacto-N-Tetraose 687.912885 1289.76172 147.76 2176.884 Lacto-N-Tetraose 424.3854625 893.4178375 125.368 1648.35178 Lacto-N-Tetraose 348.85265 767.16389 105.469 1182.654 Lacto-N-Tetraose 290.58641 641.668435 90.424 971.2455

TABLE XXII param unit DELITYP visit N mean sd median Lactodifucosyllactose mg/L UNIQUE V1 138 602.4483369 532.6734 420.450595 Lactodifucosyllactose mg/L UNIQUE V2 168 356.5481879 418.4975 226.1571229 Lactodifucosyllactose mg/L UNIQUE V3 146 276.3310824 241.1256 225.6436741 Lactodifucosyllactose mg/L UNIQUE V4 132 276.9074259 155.7537 235.3399016 Lactodifucosyllactose mg/L UNIQUE V5 127 262.4787906 105.0967 251.3244514 Lactodifucosyllactose mg/L UNIQUE V6 122 271.2121321 118.9087 251.5873565 Lactodifucosyllactose mg/L WITH V1 48 621.9134647 632.5672 446.7314741 SIBLINGS Lactodifucosyllactose mg/L WITH V2 53 324.4667808 207.8789 263.9484063 SIBLINGS Lactodifucosyllactose mg/L WITH V3 52 278.3663381 202.4931 229.1361688 SIBLINGS Lactodifucosyllactose mg/L WITH V4 50 288.8035427 153.0885 253.9644236 SIBLINGS Lactodifucosyllactose mg/L WITH V5 48 301.0084875 180.131 270.6795168 SIBLINGS Lactodifucosyllactose mg/L WITH V6 46 264.2267501 143.745 235.4535289 SIBLINGS param q1.25% q3.75% min max Lactodifucosyllactose 252.9317593 740.5602398 79.7125399 2988.924526 Lactodifucosyllactose 150.8612583 382.6521639 45.93249331 2996.415918 Lactodifucosyllactose 153.9841793 302.7349125 54.62821152 1682.713383 Lactodifucosyllactose 189.679307 314.4043267 47.58249391 1108.712256 Lactodifucosyllactose 194.6646696 328.8925958 54.61609346 659.1978832 Lactodifucosyllactose 197.9919531 321.6287901 51.57284974 738.7201617 Lactodifucosyllactose 226.8209576 724.3811897 50.73247046 3297.275118 Lactodifucosyllactose 179.9819135 365.4112845 85.89288249 1006.992878 Lactodifucosyllactose 148.2192763 307.5226659 49.85838655 1155.744731 Lactodifucosyllactose 185.9829117 369.4069428 48.51975875 755.2157228 Lactodifucosyllactose 159.9668191 374.1570922 46.25256273 823.6766534 Lactodifucosyllactose 166.4023321 323.0766056 51.0670661 789.5430687

TABLE XXIII param unit DELITYP visit N mean Sd median Sialyllacto-N- mg/L UNIQUE V1 173 120.4946707 143.5211 79.38738266 Tetraose a Sialyllacto-N- mg/L UNIQUE V2 215 120.320383 174.7271 67.4387729 Tetraose a Sialyllacto-N- mg/L UNIQUE V3 190 102.3350203 162.4454 52.40591398 Tetraose a Sialyllacto-N- mg/L UNIQUE V4 174 93.13419276 147.0611 43.84859549 Tetraose a Sialyllacto-N- mg/L UNIQUE V5 167 94.31415999 147.0746 49.15530418 Tetraose a Sialyllacto-N- mg/L UNIQUE V6 159 87.10861153 104.4478 53.16420322 Tetraose a Sialyllacto-N- mg/L WITH V1 64 131.8621806 146.2816 82.78296339 Tetraose a SIBLINGS Sialyllacto-N- mg/L WITH V2 75 156.1165221 252.4851 74.99139372 Tetraose a SIBLINGS Sialyllacto-N- mg/L WITH V3 71 120.6949772 178.2896 53.99733714 Tetraose a SIBLINGS Sialyllacto-N- mg/L WITH V4 69 107.0655282 131.3995 57.34829438 Tetraose a SIBLINGS Sialyllacto-N- mg/L WITH V5 67 116.0504387 134.8519 60.29079818 Tetraose a SIBLINGS Sialyllacto-N- mg/L WITH V6 65 104.5433758 111.7794 56.80548521 Tetraose a SIBLINGS param q1.25% q3.75% min max Sialyllacto-N- 47.18897931 124.0410768 0 837.4945131 Tetraose a Sialyllacto-N- 38.1468268 107.9080843 0 1110.361583 Tetraose a Sialyllacto-N- 27.44278706 92.1621572 0 1121.248141 Tetraose a Sialyllacto-N- 23.94542777 89.71831532 0 1187.41635 Tetraose a Sialyllacto-N- 26.76896814 90.56411096 0 1133.515897 Tetraose a Sialyllacto-N- 27.54758109 103.3993222 0 613.7754102 Tetraose a Sialyllacto-N- 51.7635368 141.5571605 0 744.0657752 Tetraose a Sialyllacto-N- 47.88950453 132.3895992 0 1819.176794 Tetraose a Sialyllacto-N- 35.99927184 100.647476 0 1092.717349 Tetraose a Sialyllacto-N- 33.00046274 108.7613672 0 619.6592317 Tetraose a Sialyllacto-N- 36.44169544 133.7701709 0 566.2366019 Tetraose a Sialyllacto-N- 36.89919403 147.2063842 6.388328416 492.4752761 Tetraose a

TABLE XXIV param unit DELITYP visit N mean Sd median Sialyllacto-N- mg/L UNIQUE V1 170 79.18246062 42.28283 68.72569099 Tetraose b Sialyllacto-N- mg/L UNIQUE V2 214 78.58361782 42.06355 70.1670586 Tetraose b Sialyllacto-N- mg/L UNIQUE V3 189 75.84894256 38.80082 67.84839427 Tetraose b Sialyllacto-N- mg/L UNIQUE V4 170 64.81255413 32.93704 58.95748683 Tetraose b Sialyllacto-N- mg/L UNIQUE V5 162 57.12345702 30.98851 50.59947014 Tetraose b Sialyllacto-N- mg/L UNIQUE V6 155 52.23655174 29.0665 44.63595583 Tetraose b Sialyllacto-N- mg/L WITH V1 64 79.88850896 33.7112 72.77711539 Tetraose b SIBLINGS Sialyllacto-N- mg/L WITH V2 75 82.99519595 33.57784 73.74143426 Tetraose b SIBLINGS Sialyllacto-N- mg/L WITH V3 71 81.41914544 34.10483 78.12830055 Tetraose b SIBLINGS Sialyllacto-N- mg/L WITH V4 67 63.55755653 31.84949 55.58607088 Tetraose b SIBLINGS Sialyllacto-N- mg/L WITH V5 66 55.35928595 31.36357 46.55150531 Tetraose b SIBLINGS Sialyllacto-N- mg/L WITH V6 63 45.79773144 24.27547 37.24666837 Tetraose b SIBLINGS param q1.25% q3.75% min max Sialyllacto-N- 53.1564539 90.75480198 14.91737174 275.7441984 Tetraose b Sialyllacto-N- 48.80673735 96.07204811 19.62994206 323.091464 Tetraose b Sialyllacto-N- 47.47623501 95.76248549 18.2993516 230.1699462 Tetraose b Sialyllacto-N- 42.03248619 77.48231245 17.91900028 203.3889653 Tetraose b Sialyllacto-N- 34.20021019 69.2258118 15.77729891 176.0331169 Tetraose b Sialyllacto-N- 31.10428435 63.33068783 14.36759859 161.4190188 Tetraose b Sialyllacto-N- 59.09832119 95.39076549 36.70322691 190.496288 Tetraose b Sialyllacto-N- 63.00766627 99.34148018 16.07949427 176.9251632 Tetraose b Sialyllacto-N- 58.42526875 97.31339889 16.14913449 192.4038113 Tetraose b Sialyllacto-N- 40.10314569 78.74882648 17.09181849 177.5190818 Tetraose b Sialyllacto-N- 31.64970623 67.96184529 17.88112005 159.9283979 Tetraose b Sialyllacto-N- 28.09090242 56.62478649 17.40002881 118.0078565 Tetraose b

Example 2

Table XXV sets out a human milk fortifier composition for in accordance with the invention. Said human milk fortifier may be for use to supplement the breast milk produced for an infant of up to 1 month of age by a primiparous mother.

TABLE XV Nutrient Per 100 kcal Energy (kcal) 100 Lipid (g) 9.76 DHA (mg) 37.26 Linoleic acid (mg) 1124.76 α-linolenic acid (mg) 107.1 ARA (mg) 47.68 ARA/DHA ratio 1.28 Linoleic/α-linolenic ratio 10.5 EPA (mg) 4.06 EPA/DHA ratio 0.11 MCT (g) 1.4 Protein (g) 0.7 Carbohydrate (g) 2.3 Minerals and electrolytes Na (mg) 71.25 K (mg) 113.62 Cl (mg) 100.12 Ca (mg) 116.41 P (mg) 69.27 Mg (mg) 8.50 Mn (μg) 7.40 Fe (mg) 2.11 Cu (mg) 0.10 Zn (mg) 1.48 I (μg) 33.76 Se (μg) 6.75 F (μg) 1.40 Cr (μg) 0.88 Mo (μg) 0.93 Vitamins and trace elements Vitamin A (μg) 518.04 Vitamin D (μg) 4.61 Vitamin E (mg) 4.3 Vitamin K (μg) 8.3 Vitamin C (mg) 24.4 Vitamin B1 (mg) 0.159 Vitamin B2 (mg) 0.227 Niacin (mg) 1.99 Vitamin B6 (mg) 0.16 Folic acid (μg) 50.17 Vitamin B12 (μg) 0.26 Pantothenic acid (mg) 1.08 Biotin (μg) 4.70 Choline (mg) 10.01 Inositol (mg) 5.59 Taurine (mg) 6.98 Carnitine (mg) 4.89 3-Fucosyllactose (mg) 13.5 Lacto-N-Fucosylpentaose II 15 (mg)

The composition according to the present invention may be formulated with many variations without departing from the scope of the invention as defined in the claims.

Example 3

Table XVI sets out an HMO human milk fortifier composition in accordance with the invention. Said human milk fortifier may be for use to supplement the breast milk produced for an infant of up to 1 month of age by a primiparous mother.

TABLE XVI HMO mg/L 3-Fucosyllactose 90 Lacto-N-Fucosylpentaose II 100 2′-Fucosyllactose 100 Lactose ??

Example 4

Table XVII sets out a human milk fortifier composition for in accordance with the invention. Said human milk fortifier may be for use to supplement the breast milk produced for an infant of up to 1 month of age by a multiparous mother. Said human milk fortifier composition may be comprised in a nutritional system with the human milk fortifier composition set out in example 2 wherein said composition of example 2 is specifally tailored for use to supplement the breast milk produced for an infant of up to 1 month of age by a primiparous mother.

TABLE XVII Nutrient Per 100 kcal Energy (kcal) 100 Lipid (g) 9.76 DHA (mg) 37.26 Linoleic acid (mg) 1124.76 α-linolenic acid (mg) 107.1 ARA (mg) 47.68 ARA/DHA ratio 1.28 Linoleic/α-linolenic ratio 10.5 EPA (mg) 4.06 EPA/DHA ratio 0.11 MCT (g) 1.4 Protein (g) 0.7 Carbohydrate (g) 2.3 Minerals and electrolytes Na (mg) 71.25 K (mg) 113.62 Cl (mg) 100.12 Ca (mg) 116.41 P (mg) 69.27 Mg (mg) 8.50 Mn (μg) 7.40 Fe (mg) 2.11 Cu (mg) 0.10 Zn (mg) 1.48 I (μg) 33.76 Se (μg) 6.75 F (μg) 1.40 Cr (μg) 0.88 Mo (μg) 0.93 Vitamins and trace elements Vitamin A (μg) 518.04 Vitamin D (μg) 4.61 Vitamin E (mg) 4.3 Vitamin K (μg) 8.3 Vitamin C (mg) 24.4 Vitamin B1 (mg) 0.159 Vitamin B2 (mg) 0.227 Niacin (mg) 1.99 Vitamin B6 (mg) 0.16 Folic acid (μg) 50.17 Vitamin B12 (μg) 0.26 Pantothenic acid (mg) 1.08 Biotin (μg) 4.70 Choline (mg) 10.01 Inositol (mg) 5.59 Taurine (mg) 6.98 Carnitine (mg) 4.89

The composition according to the present invention may be formulated with many variations without departing from the scope of the invention as defined in the claims. 

1. A human milk fortifier composition comprising one or more human milk oligosaccharide.
 2. A human milk fortifier composition according to claim 1 wherein, the composition is tailored to fortify the breast milk of a primiparous woman.
 3. A human milk fortifier composition according to claim 1 wherein, the one or more human milk oligosaccharide is selected from the group consisting of a sialylated oligosaccharide, a fucosylated oligosaccharide, an N-acetylated oligosaccharide, and combinations thereof.
 4. A human milk fortifier composition according to claim 1 wherein, the human milk oligosaccharide is selected from the group consisting of: 2′-Fucosyllactose, 3-Fucosyllactose, 6′Galactosyllactose, Blood group A Tetrasaccharide, Difucosyllacto-N-Hexose-A, Lacto-N-Difucosylhexose, Lacto-N-Fucosylpentaose-I, Lacto-N-Fucosylpentaose-II, Lacto-N-Fucosylpentaose-III, Lacto-N-Fucosylpentaose-V, Lacto-N-hexaose A, Lacto-N-hexaose B, Lacto-N-Neodifucosylhexose, Lacto-N-Neofucosylpentaose, Lacto-N-Neotetraose, Lacto-N-Tetraose, Lactodifucosyllactose, Sialyllacto-N-Tetraosce, Sialyllacto-N-Tetraose b, and combinations thereof.
 5. A human milk fortifier composition according to claim 1 wherein, the human milk oligosaccharide is selected from the group consisting of; 2′-Fucosyllactose, 3-Fucosyllactose, Lacto-N-Fucosylpentaose II, and combinations thereof.
 6. A human milk fortifier composition according to claim 1 wherein the human milk fortifier composition comprises an HMO in a range of 0.1 to 10000 mg/L.
 7. A human milk fortifier composition according to claim 1 wherein the human milk fortifier is specifically tailored for an infant of an age selected from the group consisting of, up to 4 months of age, up to 3 months of age, up to 2 months of age, up to 1 months of age, up to 2 weeks of age, and up to 1 week of age.
 8. A human milk fortifier composition according to claim 1, wherein the composition further comprises one or more ingredient selected from the group consisting of v itamins, minerals, protein, carbohydrates, and probiotics.
 9. A method of preparing a human milk fortifier tailored to fortify the breast milk of a multiparous women, comprising: measuring out an appropriate amount of a human milk fortifier comprising one or more human milk oligosaccharides: and mixing it with a diluent and/or additive. 10-14 (canceled).
 15. A method for optimising and/or preventing sub-optimal growth and development in an infant comprising administering a human milk fortifier composition comprising one or more human milk oligosaccharide to an infant in need of same. 